Identification of DNA methylation markers for human breast carcinomas using the methylation-sensitive restriction fingerprinting technique

We have developed a PCR-based method, called methylation-sensitive restriction fingerprinting (MSRF), to screen changes in DNA methylation in breast carcinomas. Two hypermethylation-containing fragments, HBC-1 (for "hypermethylation in breast cancer") and HBC-2, were identified in the ampl...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1997-03, Vol.57 (6), p.1030-1034
Main Authors: HUANG, T. H.-M, LAUX, D. E, HAMLIN, B. C, TRAN, P, TRAN, H, LUBAHN, D. B
Format: Article
Language:English
Subjects:
Base Sequence
Biological and medical sciences
Breast Neoplasms - chemistry
Breast Neoplasms - genetics
Carcinoma, Ductal, Breast - chemistry
Carcinoma, Ductal, Breast - genetics
CpG Islands
DNA Fingerprinting
DNA Methylation
DNA, Neoplasm - chemistry
DNA, Neoplasm - genetics
DNA, Neoplasm - isolation & purification
Genetic Markers
Gynecology. Andrology. Obstetrics
Humans
Mammary gland diseases
Medical sciences
Molecular Sequence Data
Polymerase Chain Reaction
Restriction Mapping
Tumors
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Summary:We have developed a PCR-based method, called methylation-sensitive restriction fingerprinting (MSRF), to screen changes in DNA methylation in breast carcinomas. Two hypermethylation-containing fragments, HBC-1 (for "hypermethylation in breast cancer") and HBC-2, were identified in the amplified breast tumor DNA relative to the amplified normal breast DNA of a patient. Nucleotide sequence analysis revealed no significant matches between the sequence of HBC-1 and the known sequences in the GenBank database, whereas the sequence of HBC-2 matched the upstream region of an antisense WT1 (Wilms' tumor suppressor gene) promoter. The methylation status in the breast tumor DNA from this patient was confirmed by Southern hybridization using HBC-1 and HBC-2 as probes, respectively. Further analysis showed that HBC-1 was methylated aberrantly in 90% (17 of 19 patients) of the primary breast carcinomas examined. This study demonstrates that MSRF provides a useful means for screening aberrant changes in DNA methylation during tumorigenesis. The commonly methylated fragments identified by MSRF could potentially supplement pathological markers currently used for cancers and additionally lead to the discovery of novel methylated tumor suppressor genes.
ISSN:0008-5472
1538-7445